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Eukaryotic Cell, June 2002, p. 401-413, Vol. 1, No. 3
1535-9778/02/$04.00+0     DOI: 10.1128/EC.1.3.401-413.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Differential Modulation of Cellular Signaling Pathways by Mild and Severe Hypovirus Strains

Todd B. Parsley, Baoshan Chen,,{dagger} Lynn M. Geletka, and Donald L. Nuss*

Center for Agricultural Biotechnology, University of Maryland Biotechnology Institute, College Park, Maryland 20742-4450

Received 7 January 2002/ Accepted 21 March 2002

Hypoviruses persistently alter multiple phenotypic traits, stably modify gene expression, and attenuate virulence (hypovirulence) of their pathogenic fungal host, the chestnut blight fungus Cryphonectria parasitica. The pleiotropic nature of these changes is consistent with hypovirus-mediated perturbation of one or more cellular signal transduction pathways. We now report that two hypoviruses that differ in the severity of symptom expression differentially perturb specific cellular signaling pathways. The C. parasitica 13-1 gene, originally identified as a hypovirus-inducible and cyclic AMP (cAMP)-regulated gene, was used to design a promoter-GFP reporter construct with which to monitor perturbation of cAMP-mediated signaling. Virus-mediated modulation of calcium/calmodulin/inositol trisphosphate-dependent signaling was monitored by measuring transcript accumulation from the C. parasitica laccase gene, lac-1. Infection by the severe hypovirus strain CHV1-EP713 caused a substantial induction of 13-1 promoter activity and a reduction of total extracellular laccase enzymatic activity (LAC-1 and LAC-3). In contrast, 13-1 promoter activity and total laccase activity were only marginally altered upon infection with the mild hypovirus strain CHV1-Euro7. However, examination of lac-1-specific transcript accumulation under previously defined culture conditions revealed that both CHV1-EP713 and CHV1-Euro7 perturbed calcium/calmodulin/inositol trisphosphate-dependent signaling. CHV1-EP713/CHV1-Euro7 chimeric viruses were used to map viral determinants responsible for modulation of cAMP-dependent signaling to domains within the central portion of the second open reading frame.

* Corresponding author. Mailing address: Center for Agricultural Biotechnology, University of Maryland Biotechnology Institute, Plant Sciences Building, Room 5115, College Park, MD 20742-4450. Phone: (301) 405-0334. Fax: (301) 314-9075. E-mail: nuss{at}umbi.umd.edu.

{dagger} Present address: Biotechnology Research Center, Guangxi University, Nanning, Guangxi 530005, People's Republic of China.

Eukaryotic Cell, June 2002, p. 401-413, Vol. 1, No. 3
1535-9778/02/$04.00+0     DOI: 10.1128/EC.1.3.401-413.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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